Skip to main content
NCBI home page
Journal of Virology logoLink to Journal of Virology
. 1994 Nov;68(11):7030–7039. doi: 10.1128/jvi.68.11.7030-7039.1994

Template-dependent, in vitro replication of rotavirus RNA.

D Chen 1, C Q Zeng 1, M J Wentz 1, M Gorziglia 1, M K Estes 1, R F Ramig 1
PMCID: PMC237140  PMID: 7933085

Abstract

A template-dependent, in vitro rotavirus RNA replication system was established. The system initiated and synthesized full-length double-stranded RNAs on rotavirus positive-sense template RNAs. Native rotavirus mRNAs or in vitro transcripts, with bona fide 3' and 5' termini, derived from rotavirus cDNAs functioned as templates. Replicase activity was associated with a subviral particle containing VP1, VP2, and VP3 and was derived from native virions or baculovirus coexpression of rotavirus genes. A cis-acting signal involved in replication was localized within the 26 3'-terminal nucleotides of a reporter template RNA. Various biochemical and biophysical parameters affecting the efficiency of replication were examined to optimize the replication system. A replication system capable of in vitro initiation has not been previously described for Reoviridae.

Full text

PDF
7030

Images in this article

7032Image
on p.7032
7033Image
on p.7033
7034Image
on p.7034
7035Image
on p.7035
7035Image
on p.7035
7036Image
on p.7036
7037Image
on p.7037
7037Image
on p.7037

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Burns J. W., Chen D., Estes M. K., Ramig R. F. Biological and immunological characterization of a simian rotavirus SA11 variant with an altered genome segment 4. Virology. 1989 Apr;169(2):427–435. doi: 10.1016/0042-6822(89)90168-2. [DOI] [PubMed] [Google Scholar]
  2. Chen D. Y., Ramig R. F. Determinants of rotavirus stability and density during CsCl purification. Virology. 1992 Jan;186(1):228–237. doi: 10.1016/0042-6822(92)90077-3. [DOI] [PubMed] [Google Scholar]
  3. Chen D., Burns J. W., Estes M. K., Ramig R. F. Phenotypes of rotavirus reassortants depend upon the recipient genetic background. Proc Natl Acad Sci U S A. 1989 May;86(10):3743–3747. doi: 10.1073/pnas.86.10.3743. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Chen D., Gombold J. L., Ramig R. F. Intracellular RNA synthesis directed by temperature-sensitive mutants of simian rotavirus SA11. Virology. 1990 Sep;178(1):143–151. doi: 10.1016/0042-6822(90)90387-7. [DOI] [PubMed] [Google Scholar]
  5. Chen D., Ramig R. F. Rescue of infectivity by in vitro transcapsidation of rotavirus single-shelled particles. Virology. 1993 Feb;192(2):422–429. doi: 10.1006/viro.1993.1057. [DOI] [PubMed] [Google Scholar]
  6. Chen D., Ramig R. F. Rescue of infectivity by sequential in vitro transcapsidation of rotavirus core particles with inner capsid and outer capsid proteins. Virology. 1993 Jun;194(2):743–751. doi: 10.1006/viro.1993.1315. [DOI] [PubMed] [Google Scholar]
  7. Cohen J., Charpilienne A., Chilmonczyk S., Estes M. K. Nucleotide sequence of bovine rotavirus gene 1 and expression of the gene product in baculovirus. Virology. 1989 Jul;171(1):131–140. doi: 10.1016/0042-6822(89)90519-9. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cohen J., Laporte J., Charpilienne A., Scherrer R. Activation of rotavirus RNA polymerase by calcium chelation. Arch Virol. 1979;60(3-4):177–186. doi: 10.1007/BF01317489. [DOI] [PubMed] [Google Scholar]
  9. Esteban R., Fujimura T., Wickner R. B. Internal and terminal cis-acting sites are necessary for in vitro replication of the L-A double-stranded RNA virus of yeast. EMBO J. 1989 Mar;8(3):947–954. doi: 10.1002/j.1460-2075.1989.tb03456.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Estes M. K., Crawford S. E., Penaranda M. E., Petrie B. L., Burns J. W., Chan W. K., Ericson B., Smith G. E., Summers M. D. Synthesis and immunogenicity of the rotavirus major capsid antigen using a baculovirus expression system. J Virol. 1987 May;61(5):1488–1494. doi: 10.1128/jvi.61.5.1488-1494.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Frilander M., Gottlieb P., Strassman J., Bamford D. H., Mindich L. Dependence of minus-strand synthesis on complete genomic packaging in the double-stranded RNA bacteriophage phi 6. J Virol. 1992 Aug;66(8):5013–5017. doi: 10.1128/jvi.66.8.5013-5017.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Fujimura T., Ribas J. C., Makhov A. M., Wickner R. B. Pol of gag-pol fusion protein required for encapsidation of viral RNA of yeast L-A virus. Nature. 1992 Oct 22;359(6397):746–749. doi: 10.1038/359746a0. [DOI] [PubMed] [Google Scholar]
  13. Fujimura T., Wickner R. B. Replicase of L-A virus-like particles of Saccharomyces cerevisiae. In vitro conversion of exogenous L-A and M1 single-stranded RNAs to double-stranded form. J Biol Chem. 1988 Jan 5;263(1):454–460. [PubMed] [Google Scholar]
  14. Gallegos C. O., Patton J. T. Characterization of rotavirus replication intermediates: a model for the assembly of single-shelled particles. Virology. 1989 Oct;172(2):616–627. doi: 10.1016/0042-6822(89)90204-3. [DOI] [PubMed] [Google Scholar]
  15. Gorziglia M. I., Collins P. L. Intracellular amplification and expression of a synthetic analog of rotavirus genomic RNA bearing a foreign marker gene: mapping cis-acting nucleotides in the 3'-noncoding region. Proc Natl Acad Sci U S A. 1992 Jul 1;89(13):5784–5788. doi: 10.1073/pnas.89.13.5784. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gottlieb P., Qiao X., Strassman J., Frilander M., Mindich L. Identification of the packaging regions within the genomic RNA segments of bacteriophage phi 6. Virology. 1994 Apr;200(1):42–47. doi: 10.1006/viro.1994.1160. [DOI] [PubMed] [Google Scholar]
  17. Gottlieb P., Strassman J., Qiao X. Y., Frucht A., Mindich L. In vitro replication, packaging, and transcription of the segmented double-stranded RNA genome of bacteriophage phi 6: studies with procapsids assembled from plasmid-encoded proteins. J Bacteriol. 1990 Oct;172(10):5774–5782. doi: 10.1128/jb.172.10.5774-5782.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Helmberger-Jones M., Patton J. T. Characterization of subviral particles in cells infected with simian rotavirus SA11. Virology. 1986 Dec;155(2):655–665. doi: 10.1016/0042-6822(86)90225-4. [DOI] [PubMed] [Google Scholar]
  19. Labbé M., Charpilienne A., Crawford S. E., Estes M. K., Cohen J. Expression of rotavirus VP2 produces empty corelike particles. J Virol. 1991 Jun;65(6):2946–2952. doi: 10.1128/jvi.65.6.2946-2952.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Liu M., Estes M. K. Nucleotide sequence of the simian rotavirus SA11 genome segment 3. Nucleic Acids Res. 1989 Oct 11;17(19):7991–7991. doi: 10.1093/nar/17.19.7991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Liu M., Mattion N. M., Estes M. K. Rotavirus VP3 expressed in insect cells possesses guanylyltransferase activity. Virology. 1992 May;188(1):77–84. doi: 10.1016/0042-6822(92)90736-9. [DOI] [PubMed] [Google Scholar]
  22. Mansell E. A., Patton J. T. Rotavirus RNA replication: VP2, but not VP6, is necessary for viral replicase activity. J Virol. 1990 Oct;64(10):4988–4996. doi: 10.1128/jvi.64.10.4988-4996.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Mason B. B., Graham D. Y., Estes M. K. In vitro transcription and translation of simian rotavirus SA11 gene products. J Virol. 1980 Mar;33(3):1111–1121. doi: 10.1128/jvi.33.3.1111-1121.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mattion N. M., Cohen J., Aponte C., Estes M. K. Characterization of an oligomerization domain and RNA-binding properties on rotavirus nonstructural protein NS34. Virology. 1992 Sep;190(1):68–83. doi: 10.1016/0042-6822(92)91193-x. [DOI] [PubMed] [Google Scholar]
  25. Minton A. P. The effect of volume occupancy upon the thermodynamic activity of proteins: some biochemical consequences. Mol Cell Biochem. 1983;55(2):119–140. doi: 10.1007/BF00673707. [DOI] [PubMed] [Google Scholar]
  26. Olkkonen V. M., Gottlieb P., Strassman J., Qiao X. Y., Bamford D. H., Mindich L. In vitro assembly of infectious nucleocapsids of bacteriophage phi 6: formation of a recombinant double-stranded RNA virus. Proc Natl Acad Sci U S A. 1990 Dec;87(23):9173–9177. doi: 10.1073/pnas.87.23.9173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Patton J. T., Gallegos C. O. Rotavirus RNA replication: single-stranded RNA extends from the replicase particle. J Gen Virol. 1990 May;71(Pt 5):1087–1094. doi: 10.1099/0022-1317-71-5-1087. [DOI] [PubMed] [Google Scholar]
  28. Patton J. T., Gallegos C. O. Structure and protein composition of the rotavirus replicase particle. Virology. 1988 Oct;166(2):358–365. doi: 10.1016/0042-6822(88)90506-5. [DOI] [PubMed] [Google Scholar]
  29. Patton J. T. Synthesis of simian rotavirus SA11 double-stranded RNA in a cell-free system. Virus Res. 1986 Dec;6(3):217–233. doi: 10.1016/0168-1702(86)90071-7. [DOI] [PubMed] [Google Scholar]
  30. Poncet D., Aponte C., Cohen J. Rotavirus protein NSP3 (NS34) is bound to the 3' end consensus sequence of viral mRNAs in infected cells. J Virol. 1993 Jun;67(6):3159–3165. doi: 10.1128/jvi.67.6.3159-3165.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Ramig R. F. Isolation and genetic characterization of temperature-sensitive mutants of simian rotavirus SA11. Virology. 1982 Jul 15;120(1):93–105. doi: 10.1016/0042-6822(82)90009-5. [DOI] [PubMed] [Google Scholar]
  32. Ramig R. F., Ward R. L. Genomic segment reassortment in rotaviruses and other reoviridae. Adv Virus Res. 1991;39:163–207. doi: 10.1016/s0065-3527(08)60795-2. [DOI] [PubMed] [Google Scholar]
  33. Wickner R. B. Double-stranded RNA virus replication and packaging. J Biol Chem. 1993 Feb 25;268(6):3797–3800. [PubMed] [Google Scholar]
  34. Zeng C. Q., Labbé M., Cohen J., Prasad B. V., Chen D., Ramig R. F., Estes M. K. Characterization of rotavirus VP2 particles. Virology. 1994 May 15;201(1):55–65. doi: 10.1006/viro.1994.1265. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Virology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES